Tailoring Fusion Proteins for Interfacial Nanoparticle Functionalization: Toward Fluorescence-Guided Imaging
Abstract Fusion proteins are specially designed molecules created by linking two or more peptides that were originally encoded by separate DNA sequences. To improve control of protein adsorption on poly(3-hydroxybutyrate) (PHB) surfaces, we developed fusion proteins comprising a polyester substrate-binding domain (SBD) fused to superfolder green fluorescent protein (sfGFP). The subunits were connected via flexible (GGGGS)n or rigid (EAAAK)n linkers with n = 1, 2, or 4. The PHB immobilization efficiency provided by these fusion proteins was extensively evaluated on microparticles and nanoparticles by using various surface-sensitive techniques. Accordingly, constructs linked by rigid sequences with n = 2 and n = 4 exhibited higher PHB adsorption capability. This suggests that reduced interdomain conformational freedom may favor configurations compatible with SBD-mediated interaction with the PHB interface. The stable association of the constructs with PHB nanoparticles enabled fluorescence-based tracking during cellular uptake experiments. The spatial orientation and polyester-binding ability conferred by such constructs via a facile, mild, and persistent association process might represent a significant biotechnological advance in the development of advanced nanosystems sensitive to fluorescence imaging.
Authors
- Maritza Araújo Todo Bom
- Guilherme Fadel Picheth (ORCID: https://orcid.org/0000-0002-2802-0180)
- Gabriela Casani Cardoso (ORCID: https://orcid.org/0000-0002-5658-7029)
- Lindomar J. C. Albuquerque (ORCID: https://orcid.org/0000-0003-1443-1795)
- Isabela Pereira Dias (ORCID: https://orcid.org/0000-0001-5139-6226)
- Camila de Almeida Salvego (ORCID: https://orcid.org/0000-0003-3573-8948)
- Ryu Masaki
- Joslaine Jacumazo (ORCID: https://orcid.org/0000-0002-5189-2833)
- Rilton Alves de Freitas (ORCID: https://orcid.org/0000-0002-7424-2999)
- Marcelo Müller‐Santos (ORCID: https://orcid.org/0000-0001-9615-6007)
- Genilza da Silva Mello
- Luiz Henrique Portela Faria (ORCID: https://orcid.org/0000-0003-3427-0482)
- Mariana Vodiani
- Edneia Amancio de Souza Ramos (ORCID: https://orcid.org/0000-0002-8855-5988)
- Emanuel Maltempi de Souza
- Giovanna Furman
- Julia Teixeira Lucena (ORCID: https://orcid.org/0009-0000-8959-0120)
- Guinther Kellerman
- Edileusa M. Gerhardt
- Maísa Colodel
- Breno C. B. Beirão
Institutions
- Universidade Tecnológica Federal do Paraná (BR)
- Brazilian Center for Research in Energy and Materials (BR)
- Universidade Federal do Paraná (BR)
Publication Details
- Journal
- ACS Omega
- Published
- 2026-10-14
- DOI
- https://doi.org/10.1021/acsomega.6c09485
- Primary Topic
- Biochemical and Structural Characterization
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Coordenação de Aperfeiçoamento de Pessoal de Nível Superior
- Conselho Nacional de Desenvolvimento Científico e Tecnológico
- Secretário de Ciência, Tecnologia e Ensino Superior, Governo do Estado de Parana
- Fundação Araucária